Todays landscape is exposed to continually increasing human activities stress. The original biotopes are disturbed and fragmented from small to smaller segments. This fact can negatively influence the biodiversity. The railway line construction is a kind of such model, in this case its planed modernisation. This study deals with a railway embankment segment nr. 190 from Hluboká n. Vlt. to Zbudov. The aim of the work was to prove the railway line as an important and valuable biotope, which needs an enhanced attention. The vegetation biodiversity on four segments was studied. Totally 182 plant species were found with no rare species. The insect (beetles) species living in the wood of old trees were studied near the railway line. There were found 21 families with 52 beetle species where 3 species were rare. These important tree old stands were mapped too. The railway line history of this segment was processed too. The railway line creates a typical biotope with high vegetation biodiversity in this segment. The presence of the rare beetles living in the wood of old trees is more important.